维甲酸诱导的人食管癌EC—109细胞分化相关基因的克隆

目的：克隆维甲酸诱导的人食管癌ＥＣ－１０９细胞分化相关基因。方法：在全反式维甲酸诱导人食管癌ＥＣ－１０９细胞系分化的基础上，采用ｍＲＮＡ差异显示技术（ｍＲＮＡｄｉｆｆｅｒｅｎｔｉａｌｄｉｓｐｌａｙ，ＤＤ）对ＥＣ－１０９细胞全反式维甲酸诱导前后的基因表达差异情况进行了分析。结果：全反式维甲酸诱导前后的细胞之间存在明显的基因表达差异，经克隆筛选获得了一些经全反式维甲酸诱导激活或抑制的差异表达基因片段。对其中一个片段（ＲＡａｌ２）的序列分析及同源性比较结果表明其与人的ａｌｐｈａ－ｃａｔｅｎｉｎ基因（Ｄ１３８６６，ＨＵＭＡＣＡ，２４３９ｂｐ）１００％同源。结论：全反式维甲酸具有调控分化相关基因表达的重要作用，在全反式维甲酸诱导前后的细胞之间存在明显的基因表达差异     

全反式维甲酸对食管癌细胞增殖及分化相关基因mRNA表达的影响

肿瘤的发生与细胞增殖与分化失调有关，诱导分化治疗是肿瘤治疗的途径之一。维甲酸类化合物是常用的诱导分化剂，对多种恶性肿瘤具有诱导分化、抑制增殖、诱导肿瘤细胞凋亡的作用。分化相关基因（NDRG1）是1997年发现的一种基因，研究表明，NDRG1在结肠腺癌、乳腺癌和前列腺癌等肿瘤组织中呈低表达，上调NDRG1表达可促进结肠上皮细胞的分化，抑制细胞增殖；为探讨全反式维甲酸（all-trans retinoic acid，ATRA）对食管癌细胞NDRG1表达及细胞增殖的影响，我们以ATRA作用于食管癌EC9706细胞，观察NDRG1表达、细胞周期及裸鼠移植瘤生长的变化。     

维甲酸诱导治疗急性早幼粒细胞白血病中细胞因子表达及其意义研…

介绍细胞因子在急性早幼粒细胞白血病的基础表达，全反式维甲酸对细胞因子分泌的调节。某些细胞因子的表达与高白细胞血症及维甲酸综合征发病机制中的关系以及细胞因子检测的临床意义。     

全反式维甲酸对早期动脉粥样硬化大鼠模型外周调节性T细胞与辅助性T细胞17平衡的影响

目的 建立早期动脉粥样硬化大鼠模型,并探讨其外周CD4+ CD25+调节性T细胞(Treg)与辅助性T细胞17(Th17)平衡变化及全反式维甲酸对平衡的影响.方法 50只8周龄SD大鼠,随机分为对照组、高脂组、免疫组、免疫高脂组以及全反式维甲酸组,对照组、免疫组予正常饲料,其他三组予高脂饲料,免疫组、免疫高脂组以及全反式维甲酸组注射卵清白蛋白进行初始免疫和强化免疫.全反式维甲酸组同时予以全反式维甲酸灌胃,16周后获取血和动脉.检测血清脂质变化,主动脉HE染色以观察动脉病变;流式细胞术检测各组大鼠外周血Treg和Th17细胞表达率,酶联免疫吸附法检测血清相关细胞因子水平.结果 成功建立早期动脉粥样硬化大鼠模型.组织形态学显示免疫高脂组内膜增生;与免疫高脂组比较,免疫组及全反式维甲酸组程度较轻,对照组及高脂组动脉完好.高脂组及免疫高脂组血清胆固醇和低密度脂蛋白较对照组、免疫组有显著增加(均P＜ 0.05),而全反式维甲酸组与其他各组比较无统计学差异.与对照组及高脂组相比,免疫组及免疫高脂组大鼠Treg细胞表达及白细胞介素10(IL-10)、转化生长因子β水平显著降低,而Th17细胞表达及IL-17、IL-6水平明显增高(均P＜0.05),全反式维甲酸组Treg表达及相关细胞因子水平与免疫高脂组比较显著升高,而Th17表达及相关因子水平明显降低(均P＜ 0.05).结论 免疫损伤结合高脂喂养可形成早期动脉粥样硬化大鼠模型,该模型存在Treg与Th17细胞平衡失调,全反式维甲酸可通过影响Th17/Treg平衡发挥抗动脉粥样硬化效应.     

全反式维甲酸对血管平滑肌细胞增生和细胞周期素依赖激酶抑制蛋白P27蛋白表达的影响

目的：探讨全反式维甲酸对细胞周期素依赖激酶抑制蛋白P27蛋白表达，血管平滑肌细胞DNA合成和增生的影响。方法：取Wistar大鼠胸主动脉血管平滑肌细胞培养，用^3H—TdR掺入率检测血管平滑肌细胞DNA合成和增生，用western—blotting印迹和图像分析方法检测细胞周期素依赖激酶抑制蛋白P27蛋白表达。结果：全反式维甲酸（2．5μmol／L）明显抑制胎牛血清（20％FBS）诱导的血管平滑肌细胞P27蛋白表达；全反式维甲酸明显抑制胎牛血清诱导的血管平滑肌细胞增生和DNA合成。结论：全反式维甲酸具有抗血管平滑肌细胞增生作用，其机制与促进P27蛋白表达有关。     

全反式维甲酸及香龙散含药血清对胃癌细胞端粒酶活性的影响

目的 分析全反式维甲酸及中药香龙散含药血清对胃癌细胞端粒酶活性的影响，并探讨端粒酶活性和细胞凋亡的关系。方法 用纯种新西兰家兔制备中药香龙散含药血清，应用TRAP分析全反式维甲酸及中药香龙散含药血清对培养的胃癌细胞端粒酶活性的影响，应用DNALadder分析及流式细胞仪观察全反式维甲酸及中药香龙散含药血清对培养的胃癌细胞凋亡的影响。结果 首先全反式维甲酸和中药香龙散含药血清对培养的胃癌细胞生长的抑制有协同作用，其次，全反式维甲酸处理胃癌细胞24小时，即可见端粒酶活性下调，中药香龙散含药血清无此作用，也不能加强全反式维甲酸对端粒酶活性下调的作用。最后，在全反式维甲酸处理的胃癌细胞保，第3天出现了细胞凋亡，并且中药香龙散含药血清对全反式维甲酸诱导的细胞凋亡有加强作用。结论 全反式维甲酸或/和中药香龙散含药血清对培养的胃癌细胞生长有抑制作用，其中全反式维甲酸可能通过抑制端粒酶活性表达而诱导了胃癌细胞凋亡，中药香龙散含药血清有加强全反式维甲酸诱导细胞凋亡的作用，其机制不清楚。     

ATRA对甲状腺鳞癌细胞株SW579细胞CDK4、CyclinD1、Rb表达的影响

目的观察全反式维甲酸（ATRA）对甲状腺鳞癌细胞株SW579细胞CDK4、CyclinD1、Rb表达的影响,以探讨其抑癌机制。方法分别以终浓度为10-7、10-6、5×10-6、10-5mol/L的ATRA作用SW579细胞株,对照组加入5μl无水乙醇,各组细胞均在培养24 h后加药。继续培养48 h,RT-PCR、Western blot检测SW579细胞CDK4、CyclinD1、Rb mRNA及其蛋白的表达。结果 ATRA作用后,RT-PCR检测结果表明,经不同浓度ATRA处理的细胞CDK4、Rb mRNA表达水平没有明显变化;CyclinD1的表达明显下调;Western blot检测结果表明经不同浓度ATRA处理的细胞CDK4蛋白表达水平没有明显变化,pRb蛋白的磷酸化水平明显下降,CyclinD1蛋白表达水平明显下降。结论 ATRA在一定浓度范围内可能通过下调甲状腺鳞癌细胞株SW579细胞CyclinD1的表达及Rb蛋白的磷酸化水平抑制细胞增殖。     

全反式维甲酸对球囊损伤大鼠胸主动脉内皮后P16及增殖细胞核抗原表达的影响

为研究全反式维甲酸对球囊损伤大鼠胸主动脉内皮后内膜增生过程、P16及增殖细胞核抗原表达的影响，球囊剥脱大鼠胸主动脉内皮，并随机将大鼠分为手术组、全反式维甲酸治疗组及对照组，各组均于术前4天灌胃至实验结束，除对照组于术后14天处死大鼠外，全反式维甲酸治疗组和手术组分别在术后2天、7天、14天和28天处死大鼠并摘除大鼠胸主动脉，通过组织学检查和免疫组织化学技术检测术后14天和28天的内膜增生情况及术后2天、7天、14天和28天P16和增殖细胞核抗原的表达及全反式维甲酸(每天30mg／kg)灌胃对它们的影响。结果发现，对照组及内皮损伤后2天均无血管内膜增厚，7天内膜开始增生，28天血管平滑肌细胞增殖减弱，但细胞外基质增加。在手术组各时间点P16的表达变化不显著，增殖细胞核抗原于球囊损伤后2天在中膜明显表达，术后7天表达达到高峰，且主要在内膜表达，14天后逐渐下降。使用全反式维甲酸治疗后，内膜增生程度及增殖细胞核抗原的表达明显降低，而P16的表达在术后2天开始升高，14天达高峰。以上结果提示，全反式维甲酸可有效抑制血管内皮损伤后内膜的增生，其机制可能与促进P16表达和抑制增殖细胞核抗原表达，从而抑制血管平滑肌细胞的增殖有关。     

全反式维甲酸对人食管癌细胞生长及survivin基因表达的影响

目的观察全反式维甲酸（ATRA）对人食管癌细胞株EC-9706的生长抑制作用以及凋亡抑制基因survivin表达的影响。方法实验组为1×10^-7、1×10^-6、1×10^-5mol／L的ATRA，空白对照组为等量RPM11640。该剂对照组为同等量RPMll640和无水乙醇。）应用MTT比色法检测细胞增殖抑制率，流式细胞仪检测细胞凋亡率，RT—PCR法测定survivin基因表达。结果ATRA作用后EC-9706细胞增殖受到抑制，呈时间和剂量依赖性，凋亡比例明显增加，survivinmRNA的表达随着作用时间的延长而呈逐渐下降趋势。结论ATRA对人食管癌细胞株EC-9706生长有抑制作用，同时可诱导其凋亡，可能与survivin基因表达下调有关。     

维甲酸诱导分化治疗甲状腺癌的临床研究

目的通过全反式维甲酸(ATRA)诱导分化治疗失分化甲状腺癌,并对其诱导分化的指标及疗效进行评价。方法15例失分化甲状腺癌,其中滤泡状癌5例,乳头状癌8例,滤泡乳头混合性癌2例。口服ATRA1~1.5mg·kg-1·d-1,持续治疗30d或60d,分别测定ATRA诱导分化治疗前后转移灶的摄碘变化、转移灶的大小及血清甲状腺球蛋白(Tg)水平,并进行比较。结果15例患者经ATRA诱导分化治疗,7例病灶的131I摄取增高;7例转移灶缩小;12例测定Tg的患者中,同时具有Tg降低、病灶摄碘增高和病灶缩小或无变化的4例,占33%。结论ATRA对失分化甲状腺癌有一定疗效,并值得进一步研究。     

Comparison of serum thyroid microsomal and thyroid peroxidase autoantibodies in thyroid diseases

Recent evidence indicates that human thyroid peroxidase (TPO) has most of the characteristics of the thyroid microsomal antigen. The question of whether TPO accounts for part or all of the antigenic activity recognized by circulating anti-microsomal antigen autoantibody (anti-M Ab) remains to be determined. The availability of an anti-TPO monoclonal antibody and of a highly purified TPO preparation allowed the development of specific and sensitive radioassays for anti-TPO autoantibody (anti-TPO Ab). In this study we compared anti-M Ab and anti-TPO Ab levels in serum from 128 subjects, including patients with Hashimoto's thyroiditis (n = 31), idiopathic myxedema (n = 11), hyperthyroid Graves' disease (n = 45), miscellaneous nonautoimmune thyroid disorders (n = 9), and normal subjects (n = 32). Anti-M Ab and anti-TPO Ab were measured by radioimmunological methods employing two different assay designs: 1) competitive radioassay (CR), based on the inhibition of radioiodinated antibody binding to human thyroid microsomes coated on microtiter wells, using a) [125I]immunoglobulin G (IgG) containing a high anti-M Ab titer (for anti-M Ab determinations), or b) [125I]anti-TPO monoclonal antibody (for anti-TPO Ab); and 2) sandwich immunoradiometric assay (IRMA) using microtiter wells coated with thyroid microsomes (for anti-M Ab determinations) or immunoaffinity-purified TPO (for anti-TPO Ab determinations) and [125I]anti-human IgG antibody. Anti-M Ab also was measured by passive hemagglutination. Anti-M Ab titers by PH closely correlated with anti-TPO Ab levels whether assayed by IRMA (r = 0.905; P less than 0.00001) or CR (r = 0.922; P less than 0.00001). Even closer correlations were found when anti-M Ab and anti-TPO Ab both were measured by the same type of radioassay procedure (IRMA, r = 0.945 and P less than 0.00001; CR, r = 0.957 and P less than 0.00001). No differences in the correlation between anti-M Ab and anti-TPO Ab results were found when the data in patients with different autoimmune thyroid disorders were analyzed separately. Further and more direct evidence for the identity of anti-M Ab and anti-TPO Ab was provided by the ability of purified TPO to completely inhibit the binding to thyroid microsomes of radioiodinated IgG preparations containing high anti-M Ab titers. In conclusion, our results provide strong support for the concept that TPO accounts for virtually all of the antigenic determinants reacting with the autoantibodies commonly termed anti-M antibodies.(ABSTRACT TRUNCATED AT 400 WORDS)     

Acromegaly and thyroid disease: prevalence of thyroid cancer

The relationship between cancer and acromegaly has been subject of study for many years. From a case of differentiated thyroid carcinoma in one of our acromegalic patients, we reviewed a series of 100 acromegalics and found two others cases of thyroid cancer, which are described in this work. From that point, we have got data from the literature about this last association and its possible pathogenesis. The prevalence of thyroid disease is increased among acromegalic patients, mainly due to nodular goiter. This association will be discussed, as well as the relation between insulin-like growth factor-I and cancer, in an effort to have a better understanding of its meaning for our cases. We concluded that it would be prudent to do periodic ultrasonographic evaluation of acromegalic patients, follow by fine needle aspiration biopsies of suspect nodules.     

Diagnosis of malignant thyroid tumors by indirect thyroid lymphography

Indirect thyroidolymphography was used for 38 patients with malignant thyroid tumors. A conclusion has been made that indirect lymphography of the thyroid is a simple and rather informative diagnostic method for malignant thyroid tumors and should be followed by spot biopsy.     

Binding of thyroid hormones to nuclear extracts of thyroid cells

Binding of T3 and T4 to soluble nuclear extracts of FRTL-5 cells, rabbit thyroid glands, and rat liver was studied. [125I]Iodo-T3 or [125I]iodo-T4 in concentration ranges of 100-fold (10-fold on each side of measured Kd) was incubated with extract at 4 C, pH 8.2, and the quantity of bound hormone was determined by collection on nitrocellulose filters. The results were corrected for nonspecific binding. Steady state (equilibrium) binding was achieved by 36 h. Apparent dissociation constants (Kd) were determined from Scatchard analysis of data pertaining to extent of binding at 36 h as a function of hormone concentration and were also calculated from kinetics of binding as the ratio of rate constants. A single class of saturable, high affinity hormone-binding sites was found. Kd values for T3 and nuclear extracts of FRTL-5 cells, rabbit thyroid gland, and rat liver were, respectively, 3.9 X 10(-11) M, 2.8 X 10(-11) M, and 4.3 X 10(-11) M from Scatchard analysis; when calculated from kinetics of hormone association, the value was 3.6 X 10(-11) M for both FRTL-5 cell and rat hepatic nuclear extract. No analysis of the time course of binding of T3 to rabbit thyroid nuclear extract was made. Kd values for T4 and FRTL-5 cell extract were 6.2 X 10(-10) M from Scatchard analysis and 5.0 X 10(-10)M from kinetic data. Half-times (t1/2) of dissociation of T3 from FRTL-5 cell and rat liver nuclear extract, calculated from association curves, were 7 and 5 h, respectively, while corresponding values determined directly and experimentally were 10.5 and 13 h. For T4 and FRTL-5 cell extract, the t1/2 of dissociation calculated from kinetics of association was 5 h; no direct experimental determination of the value was made. Numbers of T3-binding sites of FRTL-5 cell, rabbit thyroid gland, and rat liver nuclear extracts were, respectively, 71 X 10(-15), 62 X 10(-15), and 208 X 10(-15) mol/mg protein. For T4 and FRTL-5 cell extract, the value was 70 X 10(-15) mol/mg protein. The data indicate that the reaction of T3 and T4 with the various nuclear extracts can be described as reversible and bimolecular. The presence in thyroid cells of thyroid hormone nuclear binding sites suggests that they may be receptors that mediate cellular actions of these hormones within the gland itself.     

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1概述甲状腺结节系甲状腺内局部硬度和结构发生异常改变的组织肿块,是常见的内分泌疾病。它往往不是一个单独的疾病,而是多种不同疾病的临床表现。大多数甲状腺结节患者无明显症状,常由偶然触及或在体检时发现。甲状腺结节按性状可分为增生、囊肿、腺瘤、囊腺瘤等。患者的诊断需根据病史、体征、甲状腺影像学、实验室检查和甲状腺穿刺及细胞病理学等资料进行综合判断。其中,甲状腺穿刺细胞病理学检查可作为诊断的金标准。甲状腺结节中,良性结节占绝大多数(95%),其中囊性病变占25%,甲状腺癌的发生率低于5%。由于对疾病本质的认识不足,许多患…     

Influence of thyroid autoantibodies on thyroid cellular growth in vitro

Porcine thyroid follicle cells, cultured in suspension, were employed to investigate the effects of immunoglobulin preparations from patients with colloid goitre, Graves' disease or Hashimoto's thyroiditis on thyroid growth in vitro. Epidermal growth factor (EGF, 19 ng/ml) was used as a reference for maximum growth stimulation and produced a 9-fold increase in [3H]thymidine incorporation. Immunoglobulins (1000 micrograms/ml) were found to increase [3H]thymidine incorporation compared to control: from 10 normal individuals 32 +/- 4% (mean +/- SEM, % of EGF response), from 10 patients with colloid goitre 26 +/- 4% (not significantly different from normal), from 10 patients with Graves' disease 19 +/- 3% (P less than 0.05) and from 15 patients with Hashimoto's thyroiditis 11 +/- 2% (P less than 0.001). No patient immunoglobulin preparation showed activity greater than that of normal individuals. The lower growth stimulatory activity in Graves' disease and Hashimoto's thyroiditis remained after heat inactivation of serum and is thought to reflect surface binding of thyroid autoantibodies.